Article(id=1322190543883448979, tenantId=1146029695717560320, journalId=1319373162502258839, issueId=1322190415911043621, articleNumber=null, orderNo=null, doi=10.16016/j.2097-0927.202505022, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1746979200000, receivedDateStr=2025-05-12, revisedDate=1756569600000, revisedDateStr=2025-08-31, acceptedDate=null, acceptedDateStr=null, onlineDate=1793164390773, onlineDateStr=2026-10-28, pubDate=1759161600000, pubDateStr=2025-09-30, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1793164390773, onlineIssueDateStr=2026-10-28, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1793164390773, creator=13701087609, updateTime=1793164390773, updator=13701087609, issue=Issue{id=1322190415911043621, tenantId=1146029695717560320, journalId=1319373162502258839, year='2025', volume='47', issue='18', pageStart='2145', pageEnd='2288', issueExtLink='null', onlineDate='null', pubDate='1759161600000', pubDateStr='2025-09-30', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1793164360263, creator='13701087609', updateTime=1793164574764, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1322191316100960652, tenantId=1146029695717560320, journalId=1319373162502258839, issueId=1322190415911043621, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1322191316105154957, tenantId=1146029695717560320, journalId=1319373162502258839, issueId=1322190415911043621, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=2254, endPage=2261, ext={EN=ArticleExt(id=1322190544286102166, articleId=1322190543883448979, tenantId=1146029695717560320, journalId=1319373162502258839, language=EN, title=Diagnostic efficacy of metagenomic next-generation sequencing for spinal infections, columnId=1320435174995772212, journalTitle=Journal of Army Medical University, columnName=Clinical Medicine, runingTitle=null, highlight=null, articleAbstract=
Objective

To comparatively evaluate the diagnostic value of metagenomic next-generation sequencing (mNGS) versus conventional microbial culture in spinal infections.

Methods

A cross-section design was conducted on 82 consecutive patients with suspected spinal infections treated between February 2022 and January 2024 at Jiangbei Branch of First Affiliated Hospital of Army Medical University (Third Military Medical University). Microbiological culture, histopathological examination, and mNGS results from infected specimens were analyzed. Clinical diagnosis, primarily based on clinical manifestations, laboratory tests and radiologic features combined with medical history, was defined as the gold standard, and then the diagnostic performance, including sensitivity and specificity, were compared between mNGS and microbial culture.

Results

Among the 82 patients, definitive microbiological evidence was identified in 70 cases, and mNGS demonstrated a significantly higher detection rate than microbial culture (64 vs 36 cases, 78.05% vs 43.9%, P<0.05). mNGS also obtained obviously higher sensitivity, accuracy, and negative predictive value (NPV), and notably lower positive predictive value (PPV) when compared to conventional microbial culture (all P<0.05). When stratified by infection type, mNGS obtained significantly higher sensitivity and accuracy compared to microbial culture in tuberculous spinal infections (P<0.05). For non-tuberculous spinal infections, mNGS also showed superior sensitivity to microbial culture (P<0.05).

Conclusion

In patients with spinal infections, mNGS demonstrates a significantly higher pathogen detection rate than conventional microbial culture. This technique can provide early and broad-spectrum pathogenic microbiological evidence for spinal infection.

, authors=Shuang LIU1, Jinyue HE1, Hui CHEN2, Yu XIANG1, Sheng LIAO2, Zuoqiang YAN1, Huorong GOU2, Hang YANG2, Zhongrong ZHANG2, Zehua ZHANG1, 2, Jianzhong XU1, 2, authorsList=Shuang LIU, Jinyue HE, Hui CHEN, Yu XIANG, Sheng LIAO, Zuoqiang YAN, Huorong GOU, Hang YANG, Zhongrong ZHANG, Zehua ZHANG, Jianzhong XU, authorCompany=null, correspAuthors=Jianzhong XU, authorNote=null, correspAuthorsNote=null, copyrightStatement=© 2025 Journal of Army Medical University, copyrightOwner=Editorial Office of Journal of Army Medical University, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1322190546689438365, articleId=1322190543883448979, tenantId=1146029695717560320, journalId=1319373162502258839, language=CN, title=宏基因二代测序技术对脊柱感染的精准诊断效能研究, columnId=1320435175822050102, journalTitle=陆军军医大学学报, columnName=论著·临床医学, runingTitle=null, highlight=null, articleAbstract=
目的

探讨宏基因二代测序技术(metagenomic next-generation sequencing,mNGS)与常规微生物培养对脊柱感染疾病的诊断效能研究。

方法

本研究采用横断面设计,纳入2022年2月至2024年1月陆军军医大学第一附属医院江北院区入院所有诊断疑似脊柱感染患者共82例,提取感染标本的微生物培养、病理学检查和mNGS检测的结果,以临床诊断(基于病史、临床表现、实验室检查、影像学特征等综合判断)为金标准,比较mNGS与微生物培养的敏感性和特异性等诊断效能指标。

结果

82例脊柱感染患者中,得出最终病原微生物学证据的有70例,mNGS的检出率明显高于微生物培养法(64 vs 36例,78.05% vs 43.90%,P<0.05),两种方法的检出率差异有统计学意义(P<0.05)。mNGS的敏感性、准确度、阴性预测值均显著高于微生物培养(P<0.05),mNGS的阳性预测值低于微生物培养(P<0.05)。根据感染类型不同分组后,在结核性脊柱感染中,mNGS的敏感性、准确度均高于微生物培养(P<0.05)。而在非结核性脊柱感染中,mNGS的敏感性优于微生物培养(P<0.05)。

结论

在脊柱感染患者中,mNGS的检出率高于微生物培养法,该技术可为脊柱感染提供早期、广谱的病原微生物学证据。

, authors=刘爽1, 何晋月1, 陈辉2, 向渝1, 廖胜2, 闫作强1, 苟火荣2, 杨航2, 张忠荣2, 张泽华1, 2, 许建中1, 2, authorsList=刘爽, 何晋月, 陈辉, 向渝, 廖胜, 闫作强, 苟火荣, 杨航, 张忠荣, 张泽华, 许建中, authorCompany=null, correspAuthors=许建中, authorNote=null, correspAuthorsNote=
许建中,Email:
XU Jianzhong,E-mail:
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a:P<0.05

, figureFileSmall=W+RqAgWL0Y1y4867d4GKeg==, figureFileBig=NYw3iChGeyS8q7PsGIoP6A==, tableContent=null), ArticleFig(id=1322190560622916335, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1322190543883448979, language=EN, label=Table 1, caption=

Comparison between mNGS and microbiological culture in the patients [n(%)]

, figureFileSmall=null, figureFileBig=null, tableContent=
DiagnosismNGSCulturePathologyP (mNGS vs Culture)
+-+-+-
All spinal infections (n=82)64 (78.05)18 (21.95)36 (43.90)46 (56.10)63 (76.83)19 (23.17)<0.05
Tuberculous spinal infections (n=34)32 (94.12)2 (5.88)19 (55.88)15 (44.12)29 (85.29)5 (14.71)<0.05
Non-tuberculous spinal infections (n=48)32 (66.67)16 (33.33)17 (35.42)31 (64.58)34 (70.83)14 (29.17)<0.05
), ArticleFig(id=1322190561046541042, tenantId=1146029695717560320, journalId=1319373162502258839, articleId=1322190543883448979, language=CN, label=表1, caption=

mNGS与微生物培养法的检出比较 [例(%)]

, figureFileSmall=null, figureFileBig=null, tableContent=
DiagnosismNGSCulturePathologyP (mNGS vs Culture)
+-+-+-
All spinal infections (n=82)64 (78.05)18 (21.95)36 (43.90)46 (56.10)63 (76.83)19 (23.17)<0.05
Tuberculous spinal infections (n=34)32 (94.12)2 (5.88)19 (55.88)15 (44.12)29 (85.29)5 (14.71)<0.05
Non-tuberculous spinal infections (n=48)32 (66.67)16 (33.33)17 (35.42)31 (64.58)34 (70.83)14 (29.17)<0.05
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宏基因二代测序技术对脊柱感染的精准诊断效能研究
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刘爽 1 , 何晋月 1 , 陈辉 2 , 向渝 1 , 廖胜 2 , 闫作强 1 , 苟火荣 2 , 杨航 2 , 张忠荣 2 , 张泽华 1, 2 , 许建中 1, 2
陆军军医大学学报 | 论著·临床医学 2025,47(18): 2254-2261
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陆军军医大学学报 |论著·临床医学 2025 , 47 (18) : 2254 -2261
宏基因二代测序技术对脊柱感染的精准诊断效能研究
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刘爽1, 何晋月1, 陈辉2, 向渝1, 廖胜2, 闫作强1, 苟火荣2, 杨航2, 张忠荣2, 张泽华1, 2, 许建中1, 2
作者信息
  • 1陆军军医大学(第三军医大学)第一附属医院骨科,重庆
  • 2陆军军医大学(第三军医大学)第一附属医院江北院区(陆军第九五八医院)脊柱外科,重庆
通讯作者:
许建中,Email:
XU Jianzhong,E-mail:
Diagnostic efficacy of metagenomic next-generation sequencing for spinal infections
Shuang LIU1, Jinyue HE1, Hui CHEN2, Yu XIANG1, Sheng LIAO2, Zuoqiang YAN1, Huorong GOU2, Hang YANG2, Zhongrong ZHANG2, Zehua ZHANG1, 2, Jianzhong XU1, 2
Affiliations
  • 1Department of Orthopedics,First Affiliated Hospital,Army Medical University (Third Military Medical University),Chongqing
  • 2Department of Spine Surgery,Jiangbei Branch of First Affiliated Hospital of Army Medical University (Third Military Medical University),No. 958 Hospital of PLA Army,Chongqing,China
出版时间: 2025-09-30 doi: 10.16016/j.2097-0927.202505022
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目的

探讨宏基因二代测序技术(metagenomic next-generation sequencing,mNGS)与常规微生物培养对脊柱感染疾病的诊断效能研究。

方法

本研究采用横断面设计,纳入2022年2月至2024年1月陆军军医大学第一附属医院江北院区入院所有诊断疑似脊柱感染患者共82例,提取感染标本的微生物培养、病理学检查和mNGS检测的结果,以临床诊断(基于病史、临床表现、实验室检查、影像学特征等综合判断)为金标准,比较mNGS与微生物培养的敏感性和特异性等诊断效能指标。

结果

82例脊柱感染患者中,得出最终病原微生物学证据的有70例,mNGS的检出率明显高于微生物培养法(64 vs 36例,78.05% vs 43.90%,P<0.05),两种方法的检出率差异有统计学意义(P<0.05)。mNGS的敏感性、准确度、阴性预测值均显著高于微生物培养(P<0.05),mNGS的阳性预测值低于微生物培养(P<0.05)。根据感染类型不同分组后,在结核性脊柱感染中,mNGS的敏感性、准确度均高于微生物培养(P<0.05)。而在非结核性脊柱感染中,mNGS的敏感性优于微生物培养(P<0.05)。

结论

在脊柱感染患者中,mNGS的检出率高于微生物培养法,该技术可为脊柱感染提供早期、广谱的病原微生物学证据。

脊柱感染  /  宏基因二代测序  /  微生物培养  /  敏感性与特异性
Objective

To comparatively evaluate the diagnostic value of metagenomic next-generation sequencing (mNGS) versus conventional microbial culture in spinal infections.

Methods

A cross-section design was conducted on 82 consecutive patients with suspected spinal infections treated between February 2022 and January 2024 at Jiangbei Branch of First Affiliated Hospital of Army Medical University (Third Military Medical University). Microbiological culture, histopathological examination, and mNGS results from infected specimens were analyzed. Clinical diagnosis, primarily based on clinical manifestations, laboratory tests and radiologic features combined with medical history, was defined as the gold standard, and then the diagnostic performance, including sensitivity and specificity, were compared between mNGS and microbial culture.

Results

Among the 82 patients, definitive microbiological evidence was identified in 70 cases, and mNGS demonstrated a significantly higher detection rate than microbial culture (64 vs 36 cases, 78.05% vs 43.9%, P<0.05). mNGS also obtained obviously higher sensitivity, accuracy, and negative predictive value (NPV), and notably lower positive predictive value (PPV) when compared to conventional microbial culture (all P<0.05). When stratified by infection type, mNGS obtained significantly higher sensitivity and accuracy compared to microbial culture in tuberculous spinal infections (P<0.05). For non-tuberculous spinal infections, mNGS also showed superior sensitivity to microbial culture (P<0.05).

Conclusion

In patients with spinal infections, mNGS demonstrates a significantly higher pathogen detection rate than conventional microbial culture. This technique can provide early and broad-spectrum pathogenic microbiological evidence for spinal infection.

spinal infection  /  metagenomic next-generation sequencing  /  microbial culture  /  sensitivity and specificity
刘爽, 何晋月, 陈辉, 向渝, 廖胜, 闫作强, 苟火荣, 杨航, 张忠荣, 张泽华, 许建中. 宏基因二代测序技术对脊柱感染的精准诊断效能研究. 陆军军医大学学报, 2025 , 47 (18) : 2254 -2261 . DOI: 10.16016/j.2097-0927.202505022
Shuang LIU, Jinyue HE, Hui CHEN, Yu XIANG, Sheng LIAO, Zuoqiang YAN, Huorong GOU, Hang YANG, Zhongrong ZHANG, Zehua ZHANG, Jianzhong XU. Diagnostic efficacy of metagenomic next-generation sequencing for spinal infections[J]. Journal of Army Medical University, 2025 , 47 (18) : 2254 -2261 . DOI: 10.16016/j.2097-0927.202505022
脊柱感染是脊柱外科领域的重要挑战,其中结核性脊柱感染因早期临床症状隐匿(如低热、盗汗、局部疼痛)、影像学表现与化脓性感染或肿瘤重叠,导致诊断延误率高达40%[1-2]。传统诊断方法依赖病原微生物培养(如结核分枝杆菌培养需5~7周)和病理学检查,但阳性率显著受限,解放军总医院第八医学中心的研究显示,112例脊柱感染患者中,结核培养阳性率仅34.69%,而非结核感染的细菌培养阳性率也仅有53.97%[3]。此外,病理学检查对非典型感染(如布鲁氏菌、真菌)的鉴别能力不足,进一步加剧了临床诊疗的困境[4-5]。
宏基因二代测序技术(metagenomic next-generation sequencing,mNGS)能够快速、高效地对脊柱感染性疾病的病原微生物进行检测,并且对于结核性脊柱感染疾病有着较高的检出率、敏感度和特异度[6]。该技术是诊断与鉴别脊柱感染性疾病较好、较快的方法,可为结核性脊柱感染性疾病的诊断提供指导[7-8]。mNGS技术通过无偏倚检测样本中所有微生物的核酸序列,实现了病原学的快速(48 h内)和广谱(覆盖细菌、真菌、病毒及罕见病原体)诊断[9]。其革新性在于突破传统方法的“靶向检测”局限,例如在脊柱结核中,mNGS可同时检测结核分枝杆菌复合群(mycobacterium tuberculosis complex ,MTBC)与其他共感染病原体(如爱泼斯坦-巴尔病毒(Epstein-Barr virus,EBV)或曲霉菌),显著提高混合感染的检出率[10]。解放军总医院的多中心研究表明,mNGS对脊柱结核的阳性检出率达69.39%,诊断效能(AUC=0.839)显著优于微生物培养和结核分枝杆菌/利福平耐药实时荧光定量核酸扩增检测(mycobacterium tuberculosis/Rifampicin resistance real-time PCR assay,Xpert MTB/RIF)[3]。青岛大学附属医院的临床实践进一步验证,mNGS联合肺泡灌洗液标本可将诊断敏感度提升至95.02%,尤其适用于免疫功能抑制或经验性治疗无效的复杂病例[11]。
当前,mNGS技术正逐步成为脊柱感染诊断的“第二金标准”,其临床价值不仅体现在早期精准分型(结核性vs非结核性),还为个体化抗感染治疗和手术决策提供关键依据。然而,技术成本、数据解读标准化及假阳性问题仍需多学科协作优化[12]。且其与常规微生物培养对结核性与非结核性脊柱感染疾病的诊断价值对比尚不明确,本文旨在探讨二者在诊断过程中诊断效能对比,为脊柱感染临床治疗提供依据。
本研究采用横断面设计,连续纳入2022年2月至2024年1月陆军军医大学第一附属医院江北院区入院所有诊断疑似脊柱感染患者共82例。纳入标准:①经临床病史采集、体格检查、影像学评估及实验室检测综合判定为临床高度疑似脊柱感染者;②存在明确神经功能损伤;③同时完成mNGS、病原微生物培养及组织病理学检测。排除标准:①合并脊柱原发/转移性肿瘤或系统性免疫性疾病;②拒绝接受组织活检及手术治疗者;③存在其他活动性感染。结合临床治疗结果、病理检查、影像学表现、实验室指标对患者进行最终诊断。参与本研究的患者均已签署知情同意书。所有病例信息已经陆军第九五八医院伦理委员会审查(ER2025KY048)。
采集基本信息、入院时临床症状、感染部位、是否有结核病(tuberculosis,TB)史、抗生素应用情况以及是否存在其他基础疾病。此外,还记录了检测方法、抗生素及抗结核药物应用种类及时间和实验室检测数据,包括常规血液检查、红细胞沉降率(erythrocyte sedimentation rate,ESR)、C反应蛋白(C-reactive protein,CRP)和降钙素原(procalcitonin,PCT)水平。最后,患者出院后继续观察随访。按照如下核心分组标准将所有入组患者分为结核性脊柱感染组及非结核性脊柱感染组。
结核性脊柱感染组必须满足以下3项特征中的至少2项:①病原学证据方面,病灶组织/脓液培养结核阳性,或经mNGS检测结核阳性,抑或经Xpert检测结核阳性;②影像学特征方面,MRI显示多椎体受累+椎间隙相对保留+椎旁冷脓肿,或CT见椎体前缘“虫蚀样”溶骨破坏;③病理学特征为组织学显示干酪样坏死或肉芽肿形成[13]。
非结核性脊柱感染组必须满足以下3项特征的至少一项+排除结核:①病原学证据方面,病灶组织/脓液培养为非结核病原菌阳性,或经mNGS检测为非结核病原菌阳性;②影像学特征方面,MRI显示椎间隙早期破坏+环形强化、边界模糊+多为单节段病变;或CT见骨质增生硬化、骨桥形成;③病理学特征表现为中性粒细胞浸润。排除结核证据:①结核斑点试验(T cell spot test,T-SPOT)阴性或结核菌素试验(purified protein derivative test,PPD)阴性;②抗结核治疗2周无效[14]。
mNGS是一种基于高通量测序技术的无偏倚病原检测方法。其核心原理是通过对临床样本中所有微生物(细菌、真菌、病毒、寄生虫等)的核酸进行随机片段化、文库构建及深度测序,结合生物信息学分析,直接识别样本中存在的病原体基因组序列,无需依赖传统预培养步骤[15]。与传统方法[如细菌培养或聚合酶链式反应靶向测序(polymerase chain reaction targeted sequencing,PCR-TS)]相比,mNGS突破了“已知病原假设”的限制,尤其适用于检测混合感染、罕见或苛养微生物(如结核分枝杆菌复合群、布鲁氏菌等)。例如,传统结核分枝杆菌培养需耗时5~7周,阳性率不足50%,而mNGS可在48 h内完成检测,显著缩短诊断周期[16]。此外,mNGS能够检测低丰度病原体(如结核分枝杆菌复合群仅需1条严格比对序列即可判读阳性),进一步提高了诊断灵敏度[9]。
脊柱感染样本的标准化采集是确保mNGS结果可靠性的关键。采用影像学引导下(如C型臂或CT)的病灶核心穿刺活检组织、脓液或血液样本,以最大限度获取感染部位的病原体核酸,或手术获取病灶组织标本。采集的临床样本置于无菌螺旋管中严格密封,并分别开展mNGS、微生物培养、病理学等检查。
脊柱感染样本的精准处理是确保mNGS检测可靠性的核心环节。①采集后样本应立即置于无菌核酸保存液(如RNAlater)中,-80 ℃低温转运以抑制核酸降解。②前处理阶段需通过机械匀浆联合溶菌酶(针对结核分枝杆菌等厚壁菌)充分裂解微生物,随后采用皂苷裂解宿主细胞并辅以DNase消化(保留微生物DNA),可将宿主核酸占比从>99%降至<85%。③建库测序采用双端150 bp高通量模式(Illumina平台),确保覆盖低丰度病原体。④生信分析需结合临床背景过滤环境污染物(如试剂盒背景菌),并通过阈值设定(如reads≥3,基因组覆盖度≥1%)提高特异性。全程需设置阴性对照(提取试剂+无菌水)及阳性对照(已知病原体标准品),严格遵循美国临床实验室改进修正案(Clinical Laboratory Improvement Amendments,CLIA)认证实验室标准,确保结果临床可溯源性。
入组所有患者已进行病理检测,结合既往文献报道研究方法[17],以临床诊断(基于病史、临床表现、实验室检查、影像学特征等综合判断)为金标准,比较mNGS与微生物培养的敏感性和特异性等诊断效能指标。
统计病例采用例(%)表示,组间比较方法采用Fisher精确检验或χ2检验。计算mNGS和培养的特异性、敏感性、准确度、阳性预测值(positive predictive value,PPV)和阴性预测值(negative predictive value,NPV)以及相应的95%CI。采用SPSS 29.0及GraphPad Prism10软件进行统计分析,采用双侧检验,P<0.05为差异具有统计学意义。
本研究共纳入脊柱感染患者82例,其中男性49例(59.76%),女性33例(40.24%),年龄12~88(56.51±16.47)岁。根据病原学分类:非结核性脊柱感染48例(58.54%),包含化脓性脊柱炎及其他特异性感染。结核性脊柱感染34例(41.46%)。感染解剖节段分布(按国际脊柱分区标准):颈椎3例(3.66%)、胸椎11例(13.41%)、胸腰段13例(15.85%)、腰椎43例(52.44%)、腰骶段12例(14.63%),见图1。
在82例患者中,结合最终临床诊断,所得出最终病原微生物学证据的有70例,其中细菌性感染占67例(95.71%),真菌性感染3例(4.29%)。细菌性病原体主要包括:①分枝杆菌属(结核分枝杆菌)。②革兰氏阳性球菌(金黄色葡萄球菌、表皮葡萄球菌)。③革兰氏阴性杆菌(大肠埃希菌、肠炎沙门菌、霍氏肠杆菌、奇异变形杆菌、嗜麦芽窄食单胞菌)。④其他特殊菌属(布鲁氏菌、丙酸杆菌、多形普雷沃氏菌)。真菌性病原体主要包括:黄曲霉菌、烟曲霉菌、隐球菌(图2)。
脊柱感染中的总体检出比较:mNGS检测出阳性64例(78.05%),阴性18例(21.95%);微生物培养检测出阳性36例(43.90%),阴性46例(56.10%),两种方法的检出率差异具有统计学意义(P<0.05),mNGS的总体检出率明显高于微生物培养法;病理结果检测出阳性63例(76.83%),阴性19例(23.17%)。结核性脊柱感染中的检出比较:mNGS法阳性32例(94.12%);微生物培养法阳性19例(55.88%);病理结果阳性29例(85.29%)。非结核性脊柱感染中的检出比较:mNGS法阳性32例(66.67%);微生物培养法阳性17例(35.42%);病理结果阳性34例(70.83%),见表1。
mNGS和微生物培养的诊断效能比较分析结果显示,传统微生物培养于mNGS在敏感性、准确度、PPV和NPV上存在显著差异。mNGS的敏感性(87.34% vs 49.21%)、准确度(79.27% vs 55.56%)、阴性预测值显著高于微生物培养(55.56% vs 30.43%,P<0.05);微生物培养的阳性预测值略高于mNGS(86.11% vs 85.94%,P<0.05);微生物培养和mNGS在特异性方面没有显著差异,mNGS的特异性为52.63%,而微生物培养的特异性为73.68%,图3A。
将样本分为结核性脊柱感染及非结核性脊柱感染2个亚组,进一步比较诊断效果。以临床诊断(基于病史、临床表现、实验室检查、影像学特征等综合判断)为金标准,分别与2组进行对比。结果显示,二者的诊断效能指标亦有差距,在结核性脊柱感染及非结核性脊柱感染中,mNGS的敏感性在2个亚组中均高于微生物培养(96.55% vs 55.17%,79.41% vs 44.12%,P<0.05,图3B、C),在结核性脊柱感染疾病中,mNGS的准确度高于微生物培养(85.29% vs 52.94%,P<0.05,图3B)。这些也和在整个样本中的诊断性能指标表现大致相同,这表明在不同的脊柱感染病原菌中,mNGS的诊断性能均优于常规微生物培养。
脊柱感染的早期诊断存在显著挑战,临床需通过多维度评估(包括病史追溯、体征分析、实验室指标、影像学特征及微生物学证据)进行综合判断[18]。然而,传统病原学检测方法(如微生物培养)阳性率低且周期长,部分病例面临病原体无法明确的风险,导致经验性抗生素治疗比例升高及治疗窗口期延误[19]。脊柱感染性疾病因症状非特异性导致诊断延迟,且延迟时间越长(如数周至数月),神经功能损伤风险显著增加[20]。mNGS基于非选择性核酸检测技术,突破了传统微生物培养对活菌增殖的依赖,通过直接捕获样本中病原体核酸片段(包括死亡微生物残留),实现对苛养菌(如结核分枝杆菌)及厌氧菌的高效识别,将诊断周期从数周缩短至48 h以内。其广谱筛查能力可同步覆盖细菌、真菌、病毒及寄生虫等19 500余种微生物(基于NCBI RefSeq数据库),在脊柱混合感染及不明原因脊柱炎中展现出独特优势[21]。尤其对于术前已接受抗生素治疗的患者,传统培养因活菌抑制导致假阴性率升高,而mNGS通过检测核酸残留仍可维持较高的病原检出率[22]。
本研究对82例疑似脊柱感染患者采用横断面设计,多种方法综合诊断策略(mNGS、微生物培养及组织病理学联合分析),并结合治疗反应动态评估(如炎性指标、影像学改善)进行结果校正,以降低单一检测方法的局限性对诊断结论的影响。综合结果来看,mNGS的病原菌检出率、敏感性、准确度、NPV均显著高于微生物培养(P<0.05),微生物培养的PPV显著高于mNGS(P<0.05),二者在特异性方面没有显著差异,与既往相关研究得出结果相符[19]。此外,在假阳性与假阴性结果比较中,与金标准相比,mNGS与微生物培养的总体假阳性率与假阴性率对比为(47.37% vs 26.32%,12.66% vs 50.79%),可见mNGS假阳性率高于微生物培养,假阴性率低于微生物培养。mNGS假阳性率高的原因:①环境污染物,采样或运输中皮肤定植菌或试剂工程菌的核酸残留可能被检出;②人源定植微生物,呼吸道、皮肤等部位的共生菌在mNGS中显示为真阳性,但实际为定植而非致病,导致假阳性解读;③样本间交叉污染可能产生假阳性结果。mNGS假阴性率低的原因:①突破培养依赖性限制,直接检测核酸,可识别难培养病原体;②抗生素抑制作用规避,即便抗生素杀灭病原体,残留核酸仍可被检出;③广谱病原覆盖能力强,同步检测细菌、真菌、病毒及寄生虫。细分为结核性脊柱感染及非结核性脊柱感染2组后,上述诊断性能指标与总体也表现大致相同。凸显出了mNGS在脊柱感染诊断中优势,可以为更多脊柱感染患者提供准确的综合治疗方案。
从获得的病原微生物学证据来看,在82例患者中,结合最终临床诊断,所得出最终病原微生物学证据的有70例。细菌性感染占67例(95.71%),真菌性感染3例(4.29%),除结核例数较多外,大肠埃希菌、金黄色葡萄球菌、表皮葡萄球菌也是脊柱感染常见的病原体,这也与其他文献报道相符[23],且本研究中丙酸杆菌、多形普雷沃氏菌、霍氏肠杆菌、奇异变形杆菌、嗜麦芽窄食单胞菌、黄曲霉菌、烟曲霉菌、隐球菌等非常见病原微生物均由mNGS检出,体现出mNGS在广谱检测能力上的表现。此结果与既往研究得出结果也大致相符[22]。
将总体分为结核性脊柱感染与非结核性脊柱感染后,比较二者的诊断效能指标时,mNGS的敏感性在2个亚组中均高于微生物培养(96.55% vs 55.17%,79.41% vs 44.12%,P<0.05),此外,在结核性脊柱感染疾病中,mNGS的准确度高于微生物培养(85.29% vs 52.94%,P<0.05)。说明相较于微生物培养,mNGS在诊断各类型脊柱感染疾病中均有明显优势[19]。
结核性脊柱感染在骨关节结核中占比例50%~60%,作为血源性播散的典型肺外结核类型,其病理特征始于椎体前缘终板下松质骨内结核分枝杆菌定植,形成特征性干酪样坏死病灶[23-24]。其进展过程呈现三大特征:①连续性椎体破坏,通常累及相邻2~3个椎体,前柱骨质侵蚀引发椎体楔形塌陷;②椎间盘迟发性受累,相较于化脓性感染早期即出现的椎间隙高度下降,结核性病变的椎间盘破坏常滞后4~6周;③冷脓肿迁移特性,坏死物质沿椎前筋膜扩散形成椎旁脓肿,部分腰段病例可延伸至腰大肌鞘形成梭形肿块。早期精准诊断对预防后凸畸形(平均Cobb角>30°)及神经功能损害(截瘫发生率12%~18%)至关重要[25]。传统微生物培养在脊柱感染性疾病中可视为“金标准”,但因其培养周期长且其阳性率较低,使得其诊断价值不高[26]。在诊断结核性脊柱感染方面,34例患者中,mNGS和微生物培养的诊断效能指标相比,mNGS的敏感性和准确度均显著高于常规微生物培养,体现出mNGS相较于常规微生物培养在诊断结核性脊柱感染疾病中的检出优势,为早期病原学证据获取及临床个体化诊疗方案制定提供依据。结合既往文献结果[23],本研究具有一定提示意义。
本研究仍存在以下不足:①本研究采用横断面设计,存在一定选择性偏倚;②样本量不足,可能影响结果的普适性:③不同的解剖标本来源可能在采集过程中掺杂异质性干扰。未来拟通过设计多中心大样本研究,并规范标本采集方法,进一步验证mNGS在脊柱感染性疾病诊断中的临床应用价值。

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2025年第47卷第18期
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doi: 10.16016/j.2097-0927.202505022
  • 接收时间:2025-05-12
  • 首发时间:2026-10-28
  • 出版时间:2025-09-30
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  • 收稿日期:2025-05-12
  • 修回日期:2025-08-31
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    1陆军军医大学(第三军医大学)第一附属医院骨科,重庆
    2陆军军医大学(第三军医大学)第一附属医院江北院区(陆军第九五八医院)脊柱外科,重庆

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2种不同金属材料的力学参数
科
Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)
属
Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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